P
US6653456B2ExpiredUtilityPatentIndex 63

Site-specific aminoglycoside derivatives and their use in immunodiagnostic assays

Assignee: ROCHE DIAGNOSTICS CORPPriority: Jul 31, 2001Filed: Jul 31, 2001Granted: Nov 25, 2003
Est. expiryJul 31, 2021(expired)· nominal 20-yr term from priority
Inventors:GHOSHAL MITALISALAMONE SALVATORE J
C07H 5/06C07K 16/44Y10T436/13Y10S530/807G01N 33/9446Y10S435/968C07H 3/06Y10S435/81A61K 31/7008Y10S436/815Y02P20/55
63
PatentIndex Score
1
Cited by
39
References
52
Claims

Abstract

A method of making a derivatized aminoglycoside includes reacting an aminoglycoside with at least 2 equivalents of a divalent metal ion in an aprotic solvent to complex two neighboring amino group and hydroxyl group pairs; reacting the non-complexed amino groups with a protecting reagent to provide protecting groups; removing the divalent metal ion to provide two unprotected amino groups; reacting one of the unprotected amino groups with a reactive substance containing an linker, a carrier, or a label; and removing the protecting groups. This method can be used to produce novel compounds and reagents.

Claims

exact text as granted — not AI-modified
What is claimed is:  
     
       1. A method of making a derivatized aminoglycoside, comprising: 
       reacting an aminoglycoside of formula (I) with at least 2 equivalents of a divalent metal ion in an aprotic solvent to complex two neighboring amino group and hydroxyl group pairs;                    
       wherein  
       A is —CH 2 NH 2 , —CHCH 3 NH 2 , or —CHCH 3 NHCH 3 ;  
       B is —H or —OH;  
       D is —H or —OH;  
       E is —NH 2  or —OH;  
       G is —NH 2 , or —NHCH 3 ;  
       J is —H or —OH;  
       L is —H, —CH 3 , or —OH;  
       Q is —H or —CH 2 OH; and  
       Y is —H, or —C(═O)CH(OH)CH 2 CH 2 NH 2 ;  
       reacting the non-complexed amino groups with a protecting reagent to provide protecting groups;  
       removing the divalent metal ion to provide two unprotected amino groups;  
       reacting one of the unprotected amino groups with a reactive substance comprising -T; and  
       removing the protecting groups to produce a compound of formula (II)                    
       wherein  
       G′ is —NH 2 , —NHCH 3 , —NH-T, or —NCH 3 -T;  
       Y′ is —H, —C(═O)CH(OH)CH 2 CH 2 NH 2 , or -T; and  
       T is a linker, a label, or a carrier, with the proviso that either G′ or Y∝, but not both, comprises -T.  
     
     
       2. The method of  claim 1 , wherein the divalent metal ion is selected from the group consisting of Cu 2+ , Co 2+ , Ni 2+ , Zn 2+ , and Cd 2+ . 
     
     
       3. The method of  claim 1 , wherein the divalent metal ion is Zn 2+ . 
     
     
       4. The method of  claim 1 , wherein the protecting groups comprise an acyl group. 
     
     
       5. The method of  claim 1 , wherein the protecting groups comprise tert-butoxycarbonyl. 
     
     
       6. The method of  claim 1 , wherein the protecting groups comprise trifluoroacetyl. 
     
     
       7. The method of  claim 1 , wherein the removing the divalent metal ion comprises treating the divalent metal ions with ammonium hydroxide. 
     
     
       8. The method of  claim 1 , wherein the removing the divalent metal ion comprises treating the divalent metal ions with a precipitating reagent. 
     
     
       9. The method of  claim 1 , wherein the reactive substance comprising -T is an acylating agent. 
     
     
       10. The method of  claim 1 , further comprising reacting G with a protecting group after the removing the divalent metal ion. 
     
     
       11. The method of  claim 10 , wherein 
       -T is a label of formula (III)                    
     
     
       12. The method of  claim 1 , wherein 
       -T comprises a poly(amino acid).  
     
     
       13. The method of  claim 1 , wherein 
       -T comprises an active ester of formula (IV)                    
     
     
       14. The method of  claim 13 , wherein -T is a moiety of formula (V)                    
     
     
       15. The method of  claim 1 , wherein 
       -T comprises aminodextran.  
     
     
       16. The method of  claim 15 , wherein -T is a moiety of formula (VI)                    
     
     
       17. The method of  claim 1 , wherein T is a linker; and wherein the method further comprises 
       reacting the linker with a label or a carrier.  
     
     
       18. The method of  claim 17 , wherein -T comprises from 1 to 20 carbon atoms. 
     
     
       19. The method of  claim 18 , wherein -T comprises from 1 to 10 carbon atoms. 
     
     
       20. The method of  claim 18 , wherein -T comprises at least one heteroatom. 
     
     
       21. The method of  claim 18 , wherein -T comprises at least one cyclic group. 
     
     
       22. The method of  claim 17 , wherein the linker and carrier together are —C(═O)—CH 2 CH 2 —C(═O)-poly(amino acid). 
     
     
       23. The method of  claim 1 , wherein -T comprises an activating group selected from the group consisting of active ester, isocyanate, isothiocyanate, thiol, imidazolyl, maleimide, carboxylic acid, urea, and biotin. 
     
     
       24. The method of  claim 1 , wherein 
       A is —CH 2 NH 2 ;  
       B is —H;  
       D is —H;  
       E is —NH 2 ;  
       J is —OH;  
       L is —H; and  
       Q is —CH 2 OH.  
     
     
       25. The method of  claim 24 , wherein G′ is —NH 2  and Y′ is -T. 
     
     
       26. The method of  claim 24 , wherein G′ is —NH-T and Y′ is —H. 
     
     
       27. The method of  claim 1 , wherein 
       A is —CH 2 NH 2 , —CHCH 3 NH 2 , or —CHCH 3 NHCH 3 ;  
       B is —H;  
       D is —H;  
       E is —NH 2 ;  
       J is —OH;  
       L is —CH 3 ; and  
       Q is —H.  
     
     
       28. The method of  claim 27 , wherein G′ is —NHCH 3  and Y′ is -T. 
     
     
       29. The method of  claim 27 , wherein G′ is —N(CH 3 )-T and Y′ is —H. 
     
     
       30. The method of  claim 1 , further comprising reacting one of the unprotected amino groups with a compound of formula (VII) after removing the divalent metal ion                    
       wherein the aminoglycoside is kanamycin; and  
       wherein A is —CH 2 NH 2 ;  
       B is —OH  
       D is —OH;  
       E is —OH;  
       G′ is —NH-T;  
       J is —OH;  
       L is —H;  
       Q is —CH 2 OH; and  
       Y′ is —C(═O)CH(OH)CH 2 CH 2 NH 2 .  
     
     
       31. The method of  claim 1 , further comprising isolating the compound together with non-site specific aminoglycoside derivatives; 
       wherein the purity of the compound is at least 90%.  
     
     
       32. The method of  claim 31 , wherein the purity of the compound is at least 95%. 
     
     
       33. The method of  claim 31 , wherein the purity of the compound is at least 97%. 
     
     
       34. The method of  claim 31 , wherein the purity of the compound is at least 99%. 
     
     
       35. A reagent, comprising: 
       a compound of formula (II)                    
       wherein  
       A is —CH 2 NH 2 , —CHCH 3 NH 2 , or —CHCH 3 NHCH 3 ;  
       B is —H or —OH;  
       D is —H or —OH;  
       E is —NH 2  or —OH;  
       G′ is —NH 2 , —NHCH 3 , —NH-T, or —NCH 3 -T;  
       J is —H or —OH;  
       L is —H, —CH 3 , or —OH;  
       Q is —H or —CH 2 OH; and  
       Y′ is —H or -T; and  
       T is a linker, a carrier, or a label, with the proviso that either G′ or Y′, but not both, comprises -T.  
     
     
       36. The reagent of  claim 35 , wherein -T comprises a reactive group selected from the group consisting of biotin, thiols, maleimides, isocyanates, isothiocyanates, imidazolyl groups, carboxylic acids, and active esters. 
     
     
       37. The reagent of  claim 35 , wherein -T is a label of formula (III)                    
     
     
       38. The reagent of  claim 35 , wherein -T comprises a poly(amino acid). 
     
     
       39. The reagent of  claim 35 , wherein -T comprises an active ester of formula (IV)                    
     
     
       40. The reagent of  claim 35 , wherein -T comprises aminodextran. 
     
     
       41. A compound of formula (II):                    
       wherein  
       A is —CH 2 NH 2 , —CHCH 3 NH 2 , or —CHCH 3 NHCH 3 ;  
       B is —H or —OH;  
       D is —H or —OH;  
       E is —NH 2  or —OH;  
       G′ is —NH 2 , —NHCH 3 , —NH—X, or —NCH 3 —X;  
       J is —H or —OH;  
       L is —H, —CH 3 , or —OH;  
       Q is —H or —CH 2 OH; and  
       Y′ is —H, —C(═O)CH(OH)CH 2 CH 2 NH 2 , or —X; and  
       wherein —X is present in only one of G′ or Y′ and is a moiety of formula (VIII), (IX), or (X)                    
     
     
       42. The compound of  claim 41 , wherein 
       A is —CH 2 NH 2 ;  
       B is —H;  
       D is —H;  
       E is —NH 2 ;  
       G′ is —NH 2 ;  
       J is —OH;  
       L is —H;  
       Q is —CH 2 OH; and  
       Y is —X.  
     
     
       43. The compound of  claim 41 , wherein 
       A is —CH 2 NH 2 ;  
       B is —H;  
       D is —H;  
       E is —NH 2 ;  
       G′ is —NH—X;  
       J is —OH;  
       L is —H;  
       Q is —CH 2 OH; and  
       Y′ is —H.  
     
     
       44. The compound of  claim 41 , wherein 
       A is —CH 2 NH 2 , —CHCH 3 NH 2 , or —CHCH 3 NHCH 3 ;  
       B is —H;  
       D is —H;  
       E is —NH 2 ;  
       G′ is —NHCH 3 ;  
       J is —OH;  
       L is —CH 3 ;  
       Q is —H; and  
       Y′ is —X.  
     
     
       45. The compound of  claim 41 , wherein 
       A is —CH 2 NH 2 , —CHCH 3 NH 2 , or —CHCH 3 NHCH 3 ;  
       B is —H;  
       D is —H;  
       E is —NH 2 ;  
       G′ is —N(CH 3 )—X;  
       J is —OH;  
       L is —CH 3 ;  
       Q is —H; and  
       Y′ is —H.  
     
     
       46. The compound of  claim 41 , wherein 
       A is —CH 2 NH 2 ;  
       B is —OH;  
       D is —OH;  
       E is —OH;  
       G′ is —NH—X;  
       J is —OH;  
       L is —H;  
       Q is —CH 2 OH; and  
       Y′ is —C(═O)CH(OH)CH 2 CH 2 NH 2 .  
     
     
       47. A test kit for determining an aminoglycoside analyte in a sample, the kit comprising, in packaged combination, an antibody specific for the analyte and a reagent comprising the compound of  claim 41 . 
     
     
       48. A reagent, comprising: 
       a compound of formula (II)                    
       wherein  
       A is —CH 2 NH 2 , —CHCH 3 NH 2 , or —CHCH 3 NHCH 3 ;  
       B is —H or —OH;  
       D is —H or —OH;  
       E is —NH 2  or —OH;  
       G′ is —NH-T;  
       J is —H or —OH;  
       L is —H, —CH 3 , or —OH;  
       Q is —H or —CH 2 OH;  
       Y′ is —C(═O)CH(OH)CH 2 CH 2 NH 2 ; and  
       T is a linker, a carrier, or a label.  
     
     
       49. The reagent of  claim 48 , wherein -T is a label of formula (III)                    
     
     
       50. The reagent of  claim 48 , wherein -T comprises a poly(amino acid). 
     
     
       51. The reagent of  claim 48 , wherein -T comprises an active ester of formula (IV)                    
     
     
       52. The reagent of  claim 48 , wherein -T comprises aminodextran.

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